1. Field of the Invention
The present invention relates to a procedure for linking cognate pairs of VH and VL encoding sequences from a population of cells enriched in particular surface antigen markers. The linking procedure involves a multiplex molecular amplification procedure capable of linking nucleotide sequences of interest in connection with the amplification, in particular polymerase chain reaction (multiplex PCR). The method is particularly advantageous for the generation of cognate pair libraries as well as combinatorial libraries of variable region encoding sequences from immunoglobulin. The invention also relates to methods for generation of chimeric human/non-human antibodies and expression libraries generated by such methods.
2. Background Art
WO 2005/042774 (Symphogen) discloses a method for linking nucleotide sequences of interest in particular cognate pairs of VH and VL encoding sequences using a multiplex molecular procedure. The sequences of interest are preferably amplified and linked from isolated single cells following limited dilution or other cell separation techniques. The reference discloses various ways of enriching a lymphocyte containing cell population to obtain a population of plasma cells that are particularly suitable for the multiplex molecular amplification procedure.